Classical and biochemical endpoints in the evaluation of phytotoxic effects caused by the herbicide trichloroacetate

Three terrestrial plant species, oat (Avena sativa ), Chinese cabbage (Brassica campestris cv. chinensis) and lettuce (Lactuca sativa), were exposed to different concentrations of herbicide TCA (sodium trichloroacetate) in a growth test according to guideline OECD # 208. Classical (i.e. germination and biomass) and biochemical (i.e., antioxydant enzyme activities) endpoints were investigated. Germination rate decreased significantly at 3.9 mg TCA kg dry soil(-1) (for oat and lettuce) and 62.5 mg TCA kg dry soil(-1) (for Chinese cabbage). Biomass decreased significantly only at 1.9 mg TCA kg dry soil(-1) (for oat and lettuce) and 15.6 mg TCA kg dry soil(-1) (for Chinese cabbage). The activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7) and glutathione reductase (EC 1.6.4.2) increased significantly at the lowest concentration of TCA tested, i.e. 0.03 mg TCA kg dry soil(-1) (for oat and lettuce) and 0.48 mg TCA kg dry soil(-1) (for Chinese cabbage). Our results showed a ranking of sensitivity among the different endpoints for the three plant species: enzyme activities>biomass>germination rate. The increase in antioxidant enzyme activities observed in this study ensured the detoxification of increased levels of active oxygen species, and presumably prevented the plants from undergoing oxidative stress damage. Thus, the use of enzyme activities will permit the detection of early injury in plant growth testing.     

Bioaccumulation of cobalt in silkworm (Bombyx mori L.) in relation to mulberry, soil and wastewater metal concentrations

The present study was planned to evaluate Co(II) toxicity in silkworm population. The soil was irrigated using synthetic wastewater to determine the effects of pH and initial cobalt concentration in its bioaccumulation in silkworm (Bombyx mori L.) food chain. The amount of cobalt in wastewater, soil, mulberry and silkworm was determined by atomic absorption spectrophotometer (AAS) analysis. The obtained results clearly indicate that silkworm can be used as template to indicate local cobalt pollution as its body length, body weight and mortality rate was found to be strongly related to cobalt concentration. Higher the cobalt amount in mulberry leaves more the toxicity to silkworm population. At 400 mg/L Co concentration and pH 4 there was maximum deposition of Co in the soil from the synthetic effluent. However, in plants and silk worm the accumulation of Co was maximum at pH 4.5 at an initial Co concentration of 400 mg/L in the synthetic effluent. The maximum cobalt found in wastewater, soil, mulberry and silkworm was 400 ± 0.01, 273.5 ± 0.04, 42.85 ± 0.01, 36.62 ± 0.22 mg/kg, respectively.     

Soil microorganisms nitrogen transformation test for abamectin 3.6 g/L EC (w/v) in loamy sand soil

The present study was conducted to determine the nitrogen transformation test of abamectin 3.6 g/L EC. This study was conducted as per OECD Guidelines for the Testing of Chemicals OECD 216. The test item abamectin 3.6 g/L emulsifiable concentrate (EC) was applied in a loamy sand soil and incubated over a period of 28 days for nitrogen transformation test at concentrations of 3.2 μL/kg soil dry weight and 16 μL/kg soil dry weight. The concentrations tested were based on one and five times the maximum recommended field application rates of 1200 mL/ha and 6000 mL/ha of abamectin 3.6 g/L EC, respectively. The deviation in soil nitrate content determined at 28 days after application of the test item to soil compared to the control was 0.14% and ? 9.25% for the single and five times test concentrations, respectively. There is no significant variation between the treatment groups and control sample. The rate of nitrate formation between 14 and 28 days after application of the test item to soil deviate from control by 10.41% and 13.74% for 3.2 and 16 μL/kg soil dry weight, respectively. Deviations in nitrate levels and nitrate formation rates in soil treated with up to and including 16 μL/kg of test item/kg soil dry weight were < 25%, compared to control indicating no significant effect occurred in nitrogen transformation.     

Phytoextraction of Soil Cobalt Using Hyperaccumulator Plants

A greenhouse study was conducted on phytoextraction of cobalt by nickel hyperaccumulators Alyssum murale and Alyssum corsicum and by two varieties of cobalt accumulator Nyssa sylvatica compared with the nonmetal accumulator crop plant Brassica juncea. The plants were grown on Sassafras sandy loam soil (<2 mg Co and 5 mg Ni/kg dry soil), amended with 1 mmol Co/kg dry soil (58.9 mg/kg), and two Ni smelter-contaminated soils, Quarry muck with 24 mg Co and 1720 mg Ni/kg dry soil and Welland loam with 37 mg Co and 2570 mg Ni/kg dry soil. All soils were adjusted to pH 6.5 to prevent Ni phytotoxicity. Of the five plant entries tested in the study, the two Alyssum species demonstrated the most promising Co phytoextraction results. In Co-amended Sassafras soil, the maximum concentration accumulated by Alyssum murale was 1320 mg Co/kg dry weight, which was almost 60 times higher than accumulation by crop plant Brassica juncea. At a single harvest after 60 days of growth, A. murale was able to extract more than 3% of Co from Co-amended soil. As expected, both Alyssum species accumulated up to 1% Ni on dry weight basis when grown on Ni-contaminated soils.

Nyssa sylvatica showed considerable Co accumulation; foliar Co concentration in the second harvest was as high as 800 mg/kg dry weight. The first few leaves that emerged were chlorotic, both in the Co-amended soil and Ni-contaminated soils, but with growth the signs of toxicity disappeared. In the Co amended soil, Co concentration in Nyssa sylvatica leaves was 30% of that found in shoots of Alyssum species, but an order of magnitude higher than that of Brassica juncea. The leaves accumulated a higher concentration compared with the stems.

Both Alyssum species and Nyssa sylvatica offer promise for phytoextraction of Co and ⁶⁰Co from contaminated or mineralized soils.     

Seasonal variations in the body composition and bioaccumulation of heavy metals in Nile tilapia collected from drainage canals in Al-Ahsa,Saudi Arabia

The body composition of Nile tilapia (Oreochromis niloticus) collected from drainage canals in Al-Ahsa, Saudi Arabia and the concentration of four heavy metals; zinc (Zn), cadmium (Cd), cobalt (Co) and lead (Pb) in both fish muscles and the water collected from this environment were assessed across the four seasons. The body composition was found to change with the seasons, with the best body composition being recorded in autumn and winter, where higher levels of protein (17.24, 17.65%), and fat (0.58, 0.71%) and lower water content (80.15, 79.86%) respectively were noted. The concentration of heavy metals in both fish muscles and the water body also varied significantly with the seasons. In the fish muscles, the highest content of Zn (0.409 mg/kg dry weight) and Cd (4.140 mg/kg dry weight) was recorded in winter, however, the highest concentration of Co (0.318 mg/kg dry weight) and Pb (1.96 mg/kg dry weight) was observed in spring and summer respectively. On the other hand, the water samples collected in autumn showed the maximum concentration of Cd (1.385 mg/L), Co (0.762 mg/L) and Pb (0.18 mg/L) however, the maximum concentration of Zn (0.0041 mg/L) was recorded in winter. With the exception of Cd, the accumulation of the studied heavy metals in fish muscles was within the safe limits for seafood recommended by various organizations.     

Changes in chemical composition of hydrophyte leaves during adaptation to aquatic environment

Leaf chemical composition of 19 hydrophytes was studied. The content of carbon, nitrogen, nonstructural carbohydrates, organic acids, minerals, and water was determined. Hydrophytes were shown to contain less carbon (below 410 mg/g dry wt in 60% species) than terrestrial plants. Hydrophytes and terrestrial plants did not differ in the nitrogen concentration in the leaves (33 and 29 mg/g dry wt, respectively). Hydrophytes were characterized by a low content of organic acids (40–90 mg/g dry wt in 60% species) and high content of mineral compounds (90–170 mg/g dry wt in 50% species). Total amount of nonstructural carbohydrates was similar in the leaves of hydrophytes and terrestrial plants (from 120 to 190 mg/g dry wt), but the proportions of various carbohydrate fractions differed substantially. In the hydrophyte leaves, the content of soluble carbohydrates was 2.4-fold lower, whereas the content of nonstructural polysaccharides 1.2-fold higher than in terrestrial plant leaves. Two groups of correlations between parameters of leaf chemical composition were distinguished: the contents of carbon, nitrogen, and soluble sugars were positively correlated, and the negative correlation was observed between these parameters and the amounts of mineral compounds, organic acids, water, and nonstructural polysaccharides. We concluded that hydrophyte leaf chemical composition reflects a specificity of plant adaptation to aquatic environment.     

Evaluating Risks to Terrestrial Wildlife from Environmental Fluoride

Information and approaches to evaluating health risks to terrestrial wildlife from fluoride contamination in the environment are few in the literature. We use environmental field data from a phosphate ore processing site, toxicity reference values (TRVs), and bioaccumulation factors relative to site conditions to develop risk-based concentrations (RBCs) for total fluoride in terrestrial biota and soil. RBCs were derived specifically for forage that are protective of terrestrial mammalian and avian ecological receptors through multiple exposure pathways, and which can be used to evaluate site remediation or as environmental monitoring action levels. Following review of the literature, we recommend fluoride TRVs for mammalian and avian wildlife, and bioavailability factors for estimating exposures related to aerial deposition of fluoride and fluoride gases. For large ungulates, information on fluoride bioavailability from feed and soil, and related effects thresholds, are summarized from studies on grazing livestock. The resultant RBCs for fluoride in forage range from 14 to 63 mg/kg dry weight, based on no-effect and low-effect concentrations, respectively. These concentrations bracket most state, provincial, and international regulatory standards of vegetation levels for protection of livestock and wildlife.     

Polycyclic aromatic hydrocarbons and heavy metals in Kostrena coastal area

The aim of this study was to determine pollution by polycyclic aromatic hydrocarbons (PAH) and heavy metals in seawater and sediment in Kostrena coastal area, as well as their toxicity using bioluminescence based tests. Total PAH concentration in seawater ranged 1.7-155.3 ng/L. The share of carcinogenetic PAH was relatively high, ranging 22-48.3%. Nickel concentrations in seawater were beyond detection limits (< 0.1 microg/L), vanadium concentrations ranged 0.66-1.96 microg/L, chrome concentrations were beyond detection limits, and copper concentrations were also beyond detection limits or extremely low (up to 0.32 microg/L). EC50 values in seawater ranged 23.80-90.90 ng/L. Correlation between total PAH concentration and toxicity of seawater showed strong connection between them (r = 0.9579). Total PAH concentration in marine sediment ranged 58.02-1116 microg/kg dry weight (d.w.). The share of carcinogenetic PAH was extremely high ranging 10-53%. Nickel concentrations in marine sediment ranged 8-24 mg/kg d.w., vanadium concentrations ranged 24-42 mg/kg d.w., chrome concentrations ranged 11-19 mg/kg d.w., and copper concentrations ranged 7-25 mg/kg d.w. EC50 values in marine sediment ranged 818-4596 microg/kg d.w. Correlation between total PAH concentration and toxicity of marine sediment showed weak connection between them (r = 0.2590). Previous studies of seawater samples from areas of the Adriatic sea under the direct influence of oil industry did not include concentrations of heavy metals, which makes our study the first to present such comprehensive results. Our results point out the need for further evaluations and following of marine environment pollution and its consequences on living organisms and marine ecosystem in whole.     

Effects of copper, lead and zinc in soil on egg development and hatching of Folsomia candida

Effects of CaCl₂, CuCl₂, ZnCl₂ and PbCl₂ on development and hatching success of eggs of Folsomia candida (Collembola) were studied under laboratory conditions. Thousands of healthy eggs from synchronized cultures were incubated in soils treated with different concentrations of the metals. Compared with the water control, egg hatch significantly decreased when concentrations of Cu, Pb and Zn reached 400, 1600 and 800 mg/kg dry soil, respectively. Values of EC_{50 (hatching)}, calculated according to the exponential model (with 95% confidence limits in brackets), were 625 (407–875), 2361 (2064–2687) and 1763 (1548–2000) mg/kg dry soils for Cu, Pb and Zn, respectively. When Cu concentration reached 1600 mg/kg dry soil, eggs became green and the percentage of green eggs changed from 5%–20% after incubation for 2 days to 15%–30% after incubation for 4 days. At 3200 mg Cu/kg dry soil, tissues inside eggs were black and shrunken.     

The bioavailability of various selenium compounds to a marine wading bird

The uptake of dietary selenium (about 3.5 mg/kg AF dry wt) as selenomethionine, selenocystine, selenite, selenate, and fish selenium in the plasma and red blood cells (RBC) of the oystercatcher has been investigated. The birds received the various selenium compounds subsequently, for at least 9 wk. After dietary supplementation of selenocystine, selenite, and selenate, plasma selenium was about 350 μg/L and RBC selenium 2.1 mg/kg dry wt. After supplementation of selenomethionine, the plasma concentration increased to 630 μg/L, and the RBC concentration to 4.1 mg/kg dry wt. When the fodder contained 3.1 mg/kg fish Se, an average plasma and RBC concentration of 415 μg/L and 14.4 mg/kg dry wt, respectively, was measured. The maximal increase of the selenium concentration in the plasma was attained at first sampling, 14 d after a change in dietary selenium (selenomethione or fish Se); the uptake seemed to be a concentration-regulated process. RBC concentrations (γ in mg/kg dry wt) increased with time (X in d) according toY=a?be^?cX . Fifty percent of the total increase was attained within 17d, suggesting that diffusion into the RBC played a role. The selenium concentration in the plasma was positively correlated with the (fish) Se concentration in the fodder; the RBC concentration (60 d after the change in diet) was positively correlated with the plasma concentration. When the diet contained fish Se, the blood selenium concentrations of the captive birds were similar to the concentrations measured in field birds. Fish Se is a yet undetermined selenium compound. The present experiment showed that fish Se differed from selenomethionine, selenocystine, selenite, or selenate in uptake from the food and uptake in the RBC.     

Alleviation of lindane induced toxicity in testis of Swiss mice (Mus musculus) by combined treatment with vitamin C, vitamin E and alpha-lipoic acid

Mitigation of lindane induced toxicity in testis of Swiss mice by combined treatment with vitamin C, vitamin E and alpha-lipoic acid has been evaluated. Male healthy mice (40), 8-10 weeks old were randomly selected and divided into 4 groups, control (C); lindane (L); antioxidant (A) and antioxidant plus lindane (A+L). Group C animals were administered only the vehicle (olive oil); in group L lindane was administered orally at a dose of 40 mg/kg body wt.; in group A combination of antioxidants at a dose of 125 mg/kg body wt.(vitamin C: 50 mg/kg body wt., vitamin E: 50 mg/kg body wt. and alpha-lipoic acid: 25 mg/kg body wt.) was administered orally; in group A+L both antioxidants (125 mg/kg body wt.) and lindane (40 mg/kg body wt.) were administered at their respective doses. In group A+L antioxidants were administered 1 h prior to lindane administration. All treatments were continuously given for 60 days. Histopathological changes due to lindane intoxication indicated shrunken and distorted seminiferous tubules, sparse Leydig cells and blood vessels and atrophy in the tissue. The testis weight also decreased significantly. Lindane treated group showed increased lipid peroxidation, whereas glutathione, glutathione peroxidase, superoxide dismutase, catalase and protein were significantly decreased compared to control. Lindane induced damage was minimized by administration of antioxidants. Results suggest that combined pretreatment with antioxidants can alleviate the damage caused to testis by lindane.     

Selenium deficiency in the Federal Republic of Germany

A mean selenium of 123 mg/kg dry wt was observed in 195 samples of agricultural soils, and a mean of .158mg Se/kg dry wt in 304 samples of grassland soils collected at 354 sites in various regions of the Federal Republic of Germany. For grassland soil, a north/south gradient of Se concentrations was observed. In the industrialized regions of the North, higher Se levels were generally observed, the highest value of .652 mg Se/kg dry wt at a site in Northrhine-Westphalia. The mean selenium content of grass from the respective collection sites was .045 mg/kg dry wt in all regions of the FRG, a level insufficient for the maintenance of health of farm animals. The absence of a correlation between the soil-and grass-Se contents indicates that Se uptake by plants is not solely dependent on the presence of Se. Grass may be deficient in Se even if grown on Se-rich soils. Fixation of Se by acidic soils appears to be a major factor; the high Se levels in the soils of industrialized areas is not bioavailable. Based on these findings, it is concluded that locally produced feedstock must be supplemented with Se to prevent the outbreak of deficiency diseases in farm animals.     

Assessing measurable adverse changes to benthic invertebrate communities based on site-specific sediment toxicity testing and community data at the Anniston,Alabama PCB site

Laboratory toxicity tests were conducted for polychlorinated biphenyl (PCB)-containing sediment from Choccolocco Creek at the Anniston PCB Site. The objective of this investigation was to develop a site-specific PCB concentration–response relationship for invertebrate test organisms. Test results from reference locations were used to develop background-adjusted 10% (EC10*) and 20% (EC20*) effect concentrations for a range of survival, growth, and reproduction endpoints. The EC20* values ranged from 4.43 to 114 milligrams of PCBs per kilogram (mg/kg) of sediment on a dry weight basis for reproductive endpoints, 28 to 67.7 mg/kg sediment for growth, and 123 to 165 mg/kg sediment for survival. The aggregate EC20* response range for endpoints that demonstrated good test performance with reasonable ranges in control variability (<20%) and reference sample responses was 14.3 to 165 mg/kg. The PCB concentrations in sediments for a majority of the Site rarely exceed even the lowest of these thresholds. Given the protective process used to develop these thresholds, there is a high level of confidence that benthic communities are not impacted. This finding is supported by direct measurements of the benthic communities that have been living and reproducing on the Site.     

Effect of 24-epibrassinolide on growth, protein content and antioxidative defense system of Brassica juncea L. subjected to cobalt ion toxicity

Brassica juncea L. plants were subjected to cobalt (Co) ion (0, 5?×?10^?4, 10^?3, 1.5?×?10^?3 and 2?×?10^?3?M) toxicity and were sprayed with different concentrations of 24-epibrassinolide (24-EBL) (0, 10^?10, 10^?8 and 10^?6?M) at 15-day stage after sowing. They were sampled at 30 and 60?days after sowing and analyzed for growth parameters in terms of shoot length and number of leaves. Thereafter, leaves were excised and content of proteins and the activities of antioxidative enzymes (superoxide dismutase (SOD) (EC 1.15.1.1) catalase (CAT) (EC 1.11.1.6), ascorbate peroxidase (APOX) (EC 1.11.1.11), guaiacol peroxidase (POD) (EC 1.11.1.7) glutathione reductase (GR) (EC 1.6.4.2), monodehydroascorbate reductase (MDHAR) (EC 1.1.5.4) and dehydroascorbate reductase (DHAR) (EC 1.8.5.1)) were analyzed. The plants exposed to cobalt ion exhibited a significant decline in growth in terms of shoot length and number of leaves. However, foliar spray treatment with 24-EBL was able to alleviate the stress generated by cobalt ion and significantly improved the above parameters. The activities of antioxidative enzymes (SOD, CAT, POD, GR, APOX, MDHAR and DHAR) and protein content were also regulated considerably in leaves of plants treated with 24-EBL alone, 10^?8?M concentration being the most effective. The activities of antioxidative enzymes also increased in leaves of B. juncea plants by the application of cobalt ion to soil and consequently sprayed with 24-EBL. Similarly, the protein content was also regulated in leaves of B. juncea plants treated with 24-EBL as compared to untreated control plants, thereby revealing stress-protective properties of 24-EBL.     

Toxicity of arsenic (III) and (V) on plant growth, element uptake, and total amylolytic activity of mesquite (Prosopis juliflora x P. velutina)

The effects of arsenite [As(III)] and arsenate [As(V)] on the growth of roots, stems, and leaves and the uptake of arsenic (As), micro- and macronutrients, and total amylolytic activity were investigated to elucidate the phytotoxicity of As to the mesquite plant (Prosopis juliflora x P. velutina). The plant growth was evaluated by measuring the root and shoot length, and the element uptake was determined using inductively coupled plasma optical emission spectroscopy. The root and leaf elongation decreased significantly with increasing As(III) and As(V) concentrations; whereas, stem elongation remained unchanged. The As uptake increased with increasing As(III) or As(V) concentrations in the medium. Plants treated with 50 mg/L As(III) accumulated up to 920 mg/kg dry weight (d wt) in roots and 522 mg/kg d wt in leaves, while plants exposed to 50 mg/L As(V) accumulated 1980 and 210 mg/kg d wt in roots and leaves, respectively. Increasing the As(V) concentration up to 20 mg/L resulted in a decrease in the total amylolytic activity. On the contrary, total amylolytic activity in As(III)-treated plants increased with increasing As concentration up to 20 mg/L. The macro- and micronutrient concentrations changed in As-treated plants. In shoots, Mo and K were reduced but Ca was increased, while in roots Fe and Ca were increased but K was reduced. These changes reduced the size of the plants, mainly in the As(III)-treated plants; however, there were no visible sign of As toxicity.     

Factors affecting the selenium intake of people in transbaikalian Russia

The selenium concentration in foods grown and consumed and in plasma, red blood cells, and toenails of people living in the district of Chita in the transbaikalian part of Russia were studied in August 1991. Preliminary results from the area have suggested low selenium intakes and the possible occurrence of cardiomyopathy (Keshan disease) in the population. A low selenium concentration in foods grown locally was found: mean selenium concentration in wheat grains was 1, 5, and 28 μg/kg, respectively, in three villages studied, that of oats was beween 3–6 μg/kg, and of cow's milk 10–27 μg/kg dry matter. The selenium concentration of bread was considerably higher, between 87–337 μg/kg dry wt, presumably because wheat imported from the US had been used for baking. Occasional samples of pork, beef, and mutton contained between 32–318 μg selenium/kg dry wt. Low selenium concentrations were observed in samples of soil and river water. The mean plasma selenium concentration of 52 persons was 1.02 μmol/L, including 33 children and 19 adult subjects. The selenium concentrations in red blood cells and toenails were 1.95 μmol/L and 0.61 mg/kg, respectively. No symptoms of heart disease caused by selenium deficiency were observed. It is concluded that the selenium status of people was fairly good thanks to the contribution to dietary intake of imported wheat with a high selenium content. As the selenium concentration was very low in foods grown in the area, the selenium intake of the population will be reduced to a very low level if only locally produced foods are consumed.     

Heavy Metals in Antarctic Notothenioid Fish from South Bay, Livingston Island, South Shetlands (Antarctica)

The Pb, Cd, Cu, Zn, and Mn contents of the liver, spleen, muscle, bones, scales, gills, and the whole body of 3- to 7-year-old notothenioid Antarctic cod (Notothenia coriiceps, Richardson, 1844) were measured. The highest heavy metal concentrations obtained are as follows: Cd in liver, the mean value was 1.36 ± 0.19 mg/kg dry weight (wt); Pb and Zn in spleen, the mean values were 3.33 ± 0.86 and 143.97 ± 16.17 mg/kg dry wt, respectively; Cu in gills, 3.76 ± 1.16 mg/kg dry wt; and Mn in scales, 14.80 ± 4.77 mg/kg dry wt. The comparison with the data reported up to now shows that the metal concentrations varied within relative wide ranges. These first data obtained could be used as a baseline to investigate further relationships among metal contents in fish, their diet, and habitat.     

Enhanced production of phytoestrogenic isoflavones from hairy root cultures of Psoralea corylifolia L. Using elicitation and precursor feeding

The effect of biotic elicitors (yeast extract, chitosan), signaling molecule (salicylic acid), and polyamines (putrescine and spermidine) was studied with respect to isoflavones accumulation in hairy root cultures of Psoralea corylifolia L. Untreated hairy roots (control) accumulated 1.55% dry wt of daidzein and 0.19% dry wt of genistein. In precursor feeding experiment, phenylalanine at 2 mM concentration led to 1.3 fold higher production of daidzein (1.91% dry wt) and genistein (0.27% dry wt). In biotic elicitors, chitosan (2 mg/L) was found to be the most efficient elicitor to induce daidzein (2.78% dry wt) and genistein (0.279% dry wt) levels in hairy roots. Salicylic acid at 1 mM concentration stimulated the maximum accumulation of daidzein (2.2% dry wt) and genistein (0.228% dry wt) 2 days after elicitation. In case of polyamines, putrescine (50 mM) resulted in highest accumulation of daidzein (3.01% dry wt) and genistein (0.227% dry wt) after 5 days of addition. Present results indicated the effectiveness of elicitation and precursor feeding on isoflavones accumulation in hairy roots of P. corylifolia. This is the first report of elicitation on isoflavones production by hairy roots of P. corylifolia.     

Nitric oxide potentiates oligosaccharide-induced artemisinin production in Artemisia annua hairy roots

The purpose of the present study was to characterize the generation of nitric oxide （NO） in Artemisia annua roots induced by an oligosaccharide elicitor （OE） from Fusarium oxysporum mycelium and the potentiation role of NO in the elicitation of artemisinin accumulation. The OE （0.3 mg total sugar/mL） induced a rapid production of NO in cultures, which exhibited a biphasic time course, reaching the first plateau within 1.5 h and the second within 8 h of OE treatment. Artemisinin content in 20-day-old hairy roots was increased from 0.7mg/g dry wt to 1.3 mg/g dry wt by using the OE treatment for 4d. In the absence of OE, the NO donor sodium nitroprusside （SNP） at 10, 50 ~1 and 100 ~1 enhanced the growth of hairy roots, but had no effect on artemisinin synthesis, The combination of SNP with OE increased artemisinin content from 1.2 mg/g drywt to 2.2 mg/g dry wt, whereas the maximum production of artemisinin in cultures was 28.5 mg/L, a twofold increase over the OE treatment alone. The effects of SNP on the OE-induced artemisinin were suppressed strongly by the NO scavenger 2-（4- carboxyphenyl）-4,4,5,5-tetramethylimidazoline-l-oxyl-3-oxide （cPTIO）. The results suggest that NO can strongly potentiate elicitor-induced artemisinin synthesis in A. annua hairy roots.     

Pharmacological assay of Cordia verbenacea V: oral and topical anti-inflammatory activity, analgesic effect and fetus toxicity of a crude leaf extract

Cordia verbenacea D.C. (Borraginaceae) is a perennial bush plant that grows widely along the southeastern coast of Brazil. Its leaves have been used in folk medicine for their anti-ulcer, anti-inflammatory and cicatrizing activities. We have already described the anti-inflammatory properties of C. verbenacea and its low toxicity in different acute animal models. In the present study, we investigated the anti-inflammatory activity in sub-chronic animal models of a crude leaf lyophilized extract when administered by oral route or topically applied, and concomitantly, its analgesic potency and toxicity to the fetus. Topical administration of the extract inhibited nystatin-induced edema proportionally to the doses used, and this effect at a dose of 4.56 mg/kg body wt. was similar to that observed with 6.0 mg/kg body wt. of naproxen. In miconazole-induced edema, the leaf extract at a dose of 1.24 mg/kg body wt., orally administered, has a very similar effect as compared to nimezulide (2.5 mg/kg body wt.) and dexamethasone (0.2 mg/kg body wt.). At an oral dose of 2.48 mg/kg body wt. the extract showed a very low analgesic effect, and total absence of fetus toxicity at doses of less than 7.44 mg/kg body wt.